IMPROVED DOUBLE IMMUNOHISTOCHEMICAL STAINING METHOD FOR CRYOSTAT AND PARAFFIN WAX SECTIONS, COMBINING ALKALINE-PHOSPHATASE ANTIALKALINE PHOSPHATASE AND INDIRECT IMMUNOFLUORESCENCE

Aims-To develop an immunohistochemical staining method for cryostat an d paraffin wax sections so that two different antigens in the same sec tion of tissues could be detected by combining immunoenzyme and immuno fluorescence techniques.Methods-This double immunohistochemical staini ng method combines alkaline phosphatase-anti-alkaline phosphatase (APA AP) using New Fuchsin as a chromogen and indirect immunofluorescence. Results-APAAP staining for one antigen of this double immunohistochemi cal staining method was observed under bright field conditions alterna ting with immunofluorescence for another antigen under ultraviolet lig ht. The double exposed photograph of both easily identified the two si gnals within the same cell. Conclusions-This double immunohistochemica l staining method can overcome the disadvantages of any masking effect of the double immunoenzymatic methods and the background problems of double immunofluorescence method especially when applied to paraffin w ax sections. It also permits good morphological identification of the doubly stained cells which may be of crucial importance in studies on pathology specimens.